Thanate JuntadechKittisak YokthongwattanaSithichoke TangphatsornruangYun-kiam YapGerd KatzenmeierChanan AngsuthanasombatMahidol University. DivisionInstitute of Molecular BiosciencesMahidol University. Faculty of Science. Department of Biochemistry and Center for Excellence in Protein Structure and Function2015-06-162017-04-252015-06-162017-04-252015-06-162012-08Advances in Bioscience and Biotechnology. Vol.3, 2012, 362-369https://repository.li.mahidol.ac.th/handle/20.500.14594/1804Unicellular micro-alga Chlamydomonas reinhardtii has been recognized as a promising host for expressing recombinant proteins albeit its limited utility due to low levels of heterologous protein expression. Here, transcription of the 3.4-kb mosquito-larvicidal cry4Ba gene from Bacillus thuringiensis in transgenic C. reinhardtii chloroplasts under control of the promoter and 5’-untranslated region of photosynthetic psbA gene was accomplished. Inverted repeats in chloroplast genomes of the host strain with deleted endogenous psbA genes were selected as recombination targets. Two transformant lines were obtained by dual-phenotypic screening via exhibition of resistance to spectinomycin and restoration of photosynthetic activity. Stable and site-specific integration of intact cry4Ba and psbA genes into chloroplast genomes found in both transgenic lines implied homoplasmy of organelle populations. Achievement in cotranscription of cry4Ba and psbA transgenes revealed by RT-PCR and Northern blot analyses demonstrates the sufficiency of this system’s transcription machinery, offering the further innovation for insecticidal protein production.engMahidol UniversityChlamydomonas reinhardtiiChloroplast TransformationInverted RepeatsBt-cry4Ba TranscriptpsbA PromoterOpen Access articleArticle10.4236/abb.2012.34052